The shift in cellular metabolism, also called Warburg effect, is an emerging hallmark of cancer cells (CC) orchestrated by oncogenic proteins. MIF, a pleiotropic cytokine, is involved in CC proliferation and multiple aspects of carcinogenesis but little is known about its possible role in cellular metabolic activities. Using MIF knockdown (KD) technique and 1H-RMN spectroscopy, we evaluated the influence of MIF on MDA-MB-231 and MCF-7cells. MIF KD cells were obtained by lentiviral transduction. MDA-MB-231 MIF sc/KD and MCF-7 MIF sc/KD cells were scrapped and quenched using methanol. The solution containing the quenched cells was pipetted for intracellular metabolites extraction (methanol, chloroform and water extraction procedure). Only the aqueous phase was used. Each sample was reconstituted in phosphate buffer mixed with TSP. After acquisition on a Bruker Avance spectrometer at 500 MHz, NMR spectra are integrated in 0.04ppm subregion (binning). Multivariate analysis (Partial Least Square Discriminate Analysis [PLS-DA]) was performed on the integrated data (SIMCA P + , MKS Data Analytics Solutions). PLS-DA analysis of NMR data clearly separate the MDA-MB-231 from MCF-7 cells (ANOVA, p < 0.01). MDA-MB-231 cells show significant lower levels of intracellular glutamine, glutamate, alanine, valine, leucine, isoleucine, glucose, glycine and higher levels of lactate and myoinositol as compared to MCF-7 (Wilcox test, p <0.05). Although non-significant, some separations were also noticed between MCF-7 MIF sc and MCF-7 MIF KD as well as between MDA-MB-231 MIF sc and MDA-MB-231 MIF KD (ANOVA, p: NS). Glutamine levels were significantly decreased in MCF-7 and MDA-MB-231 cells under-expressing MIF (Wilcox test, p = 0,004 and p = 0,005 respectively). Our data show that MDA-MB-231 and MCF-7 cells have different metabolic profiles which suggest that MDA-MB-231 cells are more dependent on glutamine but less on lactate. The decrease of glutamine levels observed in both cells lines when MIF is under-expressed suggests that MIF could be involved in the regulation of this metabolite.

Pentamidine isethionate (PTMD) is an antiprotozoal agent used in different parasitic diseases as Human African Trypanosomiasis or Pneumocystis pneumonia. Given its side effects, numerous analogs are still under development worldwide. PTMD has been recently described having a potential activity in myotonic dystrophy (type 1). Here we present an UPLC method coupled to fluo or PDA detection for PTMD and one analog determination in rat plasma or urine. The chromatographic separation was achieved on a Acquity UPLC (R) HSS T3 analytical column using a mobile phase combining formic acid 0.1% (v/v) and acetonitrile (ACN) at a constant flow rate of 0.4 mL/min. Preliminary, an innovative mu SPE (solid phase extraction) procedure using Oasis (R) WCX sorbent was processed and gave satisfying and reproducible results in terms of extraction yields.Additionally, the methods were successfully validated using the accuracy profiles approach (beta = 95% and acceptance limits = 15%) over the ranges 2.88-287.52 ng/mL and from 143.76 ng/mL to 1.72 mu g/mL in rat plasma and urine for PTMD and for EBAB, from 4.23 to 423.39 ng/mL and from 211.69 ng/mL to 2.54 mu g/mL for plasma and urine, respectively.The validated protocols were applied to a pharmacokinetic (PK) study on rats and permitted to point out some relevant PK parameters on PTMD and its studied analog. (C) 2014 Elsevier B.V. All rights reserved.

The main curative treatment of colorectal cancer remains the surgery. However, when metastases are suspected, surgery is followed by a preventive chemotherapy using oxaliplatin which, unfortunately, may cause liver sinusoidal obstruction syndrome (SOS). Such hepatic damage is barely detected during or after chemotherapy due to a lack of effective diagnostic procedures, but liver biopsy. The primary objective of the present study was to identify potential early diagnosis biomarkers of SOS using a metabonomic approach. SOS was induced in rats by monocrotaline, a prototypical toxic substance. 1H NMR spectroscopy analysis of urine samples collected from rats treated with monocrotaline showed significant metabolic changes as compared to controls. During a first phase, cellular protective mechanisms such as an increased synthesis of GSH (reduced taurine) and the recruitment of cell osmolytes in the liver (betaine) were seen. In the second phase, the disturbance of the urea cycle (increased ornithine and urea reduction) leading to the depletion of NO, the alteration in the GSH synthesis (increased creatine and GSH precursors (glutamate, dimethylglycine and sarcosine)), and the liver necrosis (decrease taurine and increase creatine) all indicate the development of SOS.